It is currently believed that the disease(s) known as AIDS are caused by a virus, the Human Immunodeficiency Virus (HIV). But I have discovered that much of the destruction of the immune system in AIDS is actually caused by tiny, stealthy, antibiotic-resistant bacteria. The HIV virus weakens the immune system so that the stealthy bacteria can proliferate out-of-control, and the damage in AIDS is done just as much by these bacteria as by any virus. These very tiny L-form Cell-Wall-Deficient (CWD) antibiotic-resistant bacteria live within the cytoplasm of cells, including the phagocytic cells (e.g. monocytes, macrophages, lymphocytes, neutrophils and polymorphonuclear cells) of the immune system itself. Killing these stealthy bacteria stops the progression, can prevent re-occurrence, and can even prevent the initial occurrence, of the disease commonly known as AIDS.
It is currently believed that the diseases commonly known as Cancer, both malignant and benign, are caused by an unknown process of cellular proliferation. But I have discovered that the cellular proliferation in Cancer actually begins due to the action of the same tiny, stealthy, antibiotic-resistant bacteria as cause immune-system destruction in AIDS. These very tiny L-form Cell-Wall-Deficient (CWD) antibiotic-resistant bacteria live within the cytoplasm of cells, including the phagocytic cells (monocytes, macrophages, lymphocytes, neutrophils and polymorphonuclear cells) of the immune system itself. These bacteria cause the cell nucleus to release the mRNA signaling the Th1 cytokine cascade, without the need for conventional signaling via, for example, CD4+T-Lymphocytes. Some of these Cytokines and Chemokines, including, without limitation, Cellular Adhesion Molecule (CAM), create the environment which allows the cellular proliferation to start, and the cancerous growth to gain a foothold in the body. Killing these stealthy pathogens removes the environment needed to initiate and feed the cellular proliferation commonly called ‘Cancer’.
Activation and Re-activation of the Immune System
The tiny intra-phagocytic L-form bacteria which cause these diseases were first described at Lister Institute in 1934. However, they were never identified as being pathogenic, or to cause disease. For example, an excellent description is given in: KLIENEBERGER-NOBEL E. Filterable forms of bacteria. Bacteriol Rev. 1951 Jun; 15(2):77-103. Available at URL http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=440979 but in the ensuing half-century these pathogens were not identified as the cause of disease.
This inventor is the first person to have described (in detail) the mechanisms by which such organisms are indeed pathogenic, and has devised both molecular biology and clinical trials which demonstrate that they are at the heart of much chronic disease (see the the inventor's published scientific papers in “Special Considerations for Asserted Therapeutic or Pharmacological Utilities” below).
The immune system is activated by a number of mechanisms in the body. One of the most important is activation of the Type-1 Nuclear Receptors in the nucleus of phagocytic cells. This activation commences the process of transcription of DNA genes into the proteins and peptides which drive both the adaptive and the innate immune responses.
The ‘Vitamin D Receptor’ (VDR) has many biological functions in the body, acting on hormone systems as disparate, and ubiquitous as the Para-Thyroid Hormone (PTH) and the Gonadotrophic Releasing Hormone (GnRH). It is principally activated by the seco-steroid hormone called 1,25-dihydroxyvitamin-D (1,25-D).
The VDR has a primary role to play in activation of innate immunity, as it is responsible, inter alia, for transcription of Toll-Like Receptors TLR2 and TLR4, essential for recognition of the lipoproteins and lipopolysaccharides by which the immune system senses bacteria. It is also responsible for generation of CAMP (Cathelicidin Anti-Microbial Peptide).
A biochemical or chemical compound (a ‘ligand’) can bind to the ligand binding domain (LBD) of the VDR and activate the receptor, or it might deactivate the receptor by binding into a position which restricts activation by its natural hormone; 1,25-D. Both activation and deactivation (agonism and antagonism) are described in this inventor's paper: Marshall T G, Lee R E, Marshall F E: Common angiotensin receptor blockers may directly modulate the immune system via VDR, PPAR and CCR2b. Theor Biol Med Model. 2006Jan. 10;3(1):1. Available from URL http.//www.tbiomed.com/content/3/1/1
In particular, an exogenous chemical agent, such as a drug from the Sartan or Statin families, can activate or deactivate VDR in this manner (as a ligand). Such a ligand can also bind into the Peroxisome Proliferator-Activated Receptor (PPAR) and the GCR. Such activation and de-activation conditions the operation of the immune system, and is effective in therapy and/or prevention of Cancers and AIDS. Table 1 shows the molecular affinities exhibited by the common Sartans and Statins for several Nuclear Receptors active in the immune system.
Note that Table 1 shows considerable difference in affinity between the various Sartans and Statins licensed by the FDA as safe for therapeutic use in their primary indications. No direct equivalence can be drawn between the members in each family of drugs. Further, it is not obvious from Table 1 whether a drug is agonistic or antagonistic to receptor activation, considerable theoretical and geometric analysis is required to deduce this. This inventor has reported that Simvastatin and Olmsartan Medoxomil are both partial agonists of the VDR—see, for example: Marshall T G . “Are Statins Analogues of Vitamin D?”Letter to the Editor, The Lancet, in press.
However, even though Simvastatin is a partial VDR agonist it is not as suitable as Olmesartan Medoxomil in treatment and/or prevention of Cancers and AIDS, as it does not affect other immune-system receptors in the same beneficial manner as Olmesartan Medoxomil. But it is better than nothing, as it does partly activate innate immunity.
Weakening the Pathogens.
Such chemical compounds (ligands) can independently, and/or in addition, bind into the ligand binding domain of the G-Protein Coupled Receptors (GPCRs) located in the genomes of pathogens, including bacterial, archaeal and protozoal pathogens, including the pathogens which cause morbidity in MRSA and Tuberculosis; as well as in Autoimmune Disease, Th1 immune dysfunction, Cancers and HIV/AIDS. When such prokaryotic GPCRs are blocked by ligands, the pathogenic organism cannot function correctly, and the pathogen eventually dies. Thus the blockade of the pathogen's GPCRs and NRs gives such a ligand antimicrobial and/or anti-bacterial and/or antibiotic properties. Common Sartans, and some ‘Statins,’ already approved by the FDA for moderate hypertension and hyperlipidemia, are just a few of the compounds which can bind into the pathogens' GPCRs and NRs. By careful selection of the Statin, Sartan, or ligand, to be used in prevention and treatment of Cancers and/or AIDS, compound can be selected which additionally have action directly against the pathogenic genomes.
Examples of such direct action of these drugs on Pathogenic genomes are shown in FIG. 1 (Olmesartan Medoxomil in MRSA SAR0276), FIG. 2 (Olmesartan Medoxomil in bb0006 from Borrelia burgdorferl), FIG. 3 (Olmesartan Medoxomil in rp630 from species Rickettsia), FIG. 4 (Olmesartan Medoxomil in E.coli ydgG), FIG. 5, Olmesartan Medoxomil in Mycobacterium tuberculosis mt1133), and FIG. 7 (Simvastatin in MRSA SAR0276).
This invention additionally provides means to kill the stealthy L-form bacteria by reducing their ability to transcribe DNA genes into proteins with their 70S Ribosome. The 30S and 50S subunits of the bacterial ribosome are targeted both individually and collectively by the antibacterial agents optionally used as part of this invention. Further, this invention reduces the availability of Angiotensin II to the Angiotensin receptors on the phagocytic outer membrane, which decreases the amount of Nuclear-Factor-kappa-B produced to fuel the transcription of Cytokines and Chemokines by the activated phagocyte.
Optimally Effective Antibiotics and Dosing Regimes
These stealthy intra-cellular bacteria are so very small they have shed parts of their physical structure (their Cell Walls), and possibly also some of their plasmids, in the transformation into the L-form pathogens. Therefore, analysis of the actions of antibiotics on these organisms can only be performed after analysis of the bacterial genome, just as was performed to obtain the analysis shown in FIGS. 1, 2, 3, 4, 5, and 7.
These L-form bacteria are very, very, difficult to culture, and conventional antibiotic sensitivity testing offers little or no help in understanding the effectiveness of this invention.
Standard antibiotic regimes do not kill these intra-phagocytic bacteria. They may therefore be thought of as “antibiotic-resistant” bacteria. As L-form bacteria they are not susceptible to antibiotics in most common use, the ‘bactericidal’ antibiotics. See, for example:                Dienes L, et al: The Transformation of Typhoid Bacilli into L Forms under Various Conditions. J Bacteriol. 1950 June; 59(6): 755-764. PMID: 15436450        Dienes L: The Isolation of L Type Cultures from Bacteroides with the Aid of Penicillin and Their Reversion into the Usual Bacilli. J Bacteriol. 1948 Oct; 56(4): 445-456. PMID:1 6561 593        
This invention can kill these antibiotic-resistant bacteria. One problem is that as the bacteria are killed they release endotoxins, and/or other toxic biochemicals, into the cytoplasm, causing further disease symptoms, sometimes of even higher intensity than during the actual activity of the Th1 disease itself. This can be likened to the Jarisch-Herxheimer Reaction which has been documented when killing bacterial pathogens, most notably when killing the Treponema pallidum which are believed to cause Syphilis.
This invention solves the problem of Herxheimer-induced anaphylaxis by targeting the bacterial genome—carefully controlling the pathogen's environment, as well as the antibiotic selection and dosing regimens.
Even if the patient is given a conventional dosage of the same antibiotics described in this invention, the antibiotics may fail to totally kill the pathogens, and if they do, there is the risk of a cytokine release sufficiently intense to cause life-threatening cardiac bradycardia or life-threatening pulmonary insufficiency, both of which were observed during experimentation with this invention.
This inventor has previously performed research on novel drug-dosing regimes in several diseases. In Diabetes I explored increased efficacy and reduced side-effects from a continuous infusion of Insulin, and in Cryptorchidism and Infertility I explored increased efficacy of pulsatile dosing of the hormones LH-RH and Gn-RH.
One of the best ways to administer a continuous concentration of any drug is by using an infusion pump, like the transcutaneous infusion pump for Insulin I invented in 1982: Marshall T G , Mekhiel N, Jackman W S, Perlman K, Albiser A M: New microprocessor-based insulin controller. IEEE Trans Biomed Eng. 1983 Nov.30(11):689-95.
During my Doctoral Research we also explored pulsatile administration of drugs, and my research group was able to cure Cryptorchidism and Infertility by using pulsatile injections of hormone, rather than using a continuous concentration: Keogh E J, MacKellar A, Mallal S A, Dunn A G, McColm S C, Somerville C P, Glatthaar C, Marshall T, Attikiouzel J: Treatment of cryptorchidism with pulsatile luteinizing hormone-releasing hormone(LH-RH). J Pediatr Surg. 1983 Jun; 18(3):282-3.
It is typically believed that antibiotics administered at doses below the Minimum Inhibitory Concentration (MIC) are ineffective, and are likely to encourage the formation of antibiotic-resistant forms of the bacteria. However, when killing the antibiotic-resistant intra-cellular bacterial L-forms which cause AIDS and Cancer, I have discovered that antibiotics blocking bacterial protein synthesis by inhibiting the function of the 70S bacterial Ribosome are needed, and I have moreover discovered they are often optimally effective when delivered in a pulsatile fashion, wherein the peak concentration in the bloodstream may or may not be in excess of the MIC, but where the antibiotic concentration is allowed to decay away to a lower value before the next dose of antibiotic is given. It should be noted that the simplified pharmacokinetic model which is usually used to describe antibiotic absorption, anticipates an exponential rise of concentration to the peak value, and then a single exponential decay of that concentration (which is considered to be distributed within the plasma compartment). A pseudo-continuous concentration in the bloodstream can be achieved by dosing the drug at sufficient frequency that the next dose is absorbed before significant exponential decay from the previous dose.
The 70S Bacterial Ribosome
These L-form bacteria synthesize proteins (which they need for their survival) within a structure called a Ribosome. The bacterial ribosome is termed a ‘70S Ribosome’ and it is conventionally divided into two subunits called the ‘30S’ and the ‘50S’ subunits.
The function of the 30S subunit is primarily determined by the 16S RNA of which it is primarily comprised, while the 50S subunit's function is primarily determined by the bacterial 23S RNA. Both subunit structures are completed by a variety of proteins and additional smaller RNA elements.
Antibiotics which inhibit the 30S subunit typically bind in the region near the helix which ‘advances’ during the transcription of bacterial mRNA to bacterial proteins. Antibiotics which inhibit the 50S subunit typically bind in the region where the tRNA docks, or in the region termed the Peptidyl Transferase Center (PTC), or in the region where the partially assembled protein travels through the body of the 50S subunit, prior to emerging as a completed protein from the ribosome.
Antibiotics which act by inhibiting actions of the 30S ribosomal subunit include Minocycline, Minocycline hydrochloride, Demeclocycline, Demeclocycline hydrochloride, Tigecycline, Tetracycline, Oxytetracycline, Doxycycline, Doxycycline hyclate, Spectinomycin, Hygromycin, Paromomycin, Streptomycin, Kanamycin, Gentamicin, Tobramycin, Amakacin, Netilmicin, Neomycin,
Antibiotics which act by inhibiting actions of the 50S ribosomal subunit include Azithromycin, Clarithromycin, Clindamycin, Chloramphenicol, Linezolid, Erythromycin, Roxithromycin, Troleandomycin, Tylocin, Sparsomycin, Carbomycin A, Sparsomycin, Lincomycin, Cethromycin, Telithromycin, Tiamulin, Dalfopristin and Quinupristin.
The effectiveness of this invention is partially due to its control of the bacterial environment, and partially due to the use of antibiotics which act symbiotically on different areas of the ribosome, reducing the statistical likelihood that any bacteria species will have developed resistance mechanisms which simultaneously overcome all the methods being used by this invention to weaken protein synthesis by the ribosome.
The Vitamin D metabolites in Th1 Inflammation.
Th1 inflammation is customarily defined as inflammation which produces an inflammatory cytokine profile including significant ‘Interferon-gamma.’ Moreover, since this Th1 cytokine release also catalyzes the synthesis of the secosteroid hormone 1,25-dihydroxyvitamin-D (1,25-D) in the infected phagocytes, it is often possible to measure the proportion of 1,25-D which leaches into the bloodstream, together with plasma 25-hydroxyvitamin-D (25-D), and estimate the extent of Thi process in well-perfused, inflamed tissue.
One estimate of Th1 inflammation is performed by calculating the D-Ratio, the ratio of 1,25-D (in pg/mi) to the 25-D (in ng/ml). The value for a healthy population is 1.25, and this ratio is often elevated in Th1 immune disease because 25-D is down-regulated, and energetically converted to 1,25-D in the cytoplasm of the phagocytes and leukocytes. The conversion of 25-D to 1,25-D is catalyzed by the action of the cytokine Interferon-gamma. The measurement and prediction based on serum-based assays of the D-metabolites is only valid if the patient is not taking any supplements containing Vitamin D, and where the value of the presenting 25-D assay is between 14 ng/ml and 20 ng/ml.
This inventor has discovered that levels of 25-D higher than 25 ng/ml are associated with that metabolite exerting immunosuppression. It reduces innate immune activity by displacing 1,25-D from the ligand binding domain of the VDR and de-activating that receptor, preventing the transcription of genes essential to proper functioning of innate immunity.
Many investigators have noted that the level of 25-D falls below 20 ng/ml in patients with the Th1 inflammatory diseases, but that observation has not heretofore been recognized as a useful marker for the disease process itself. It has been mistakenly linked with an aberrant calcium metabolism. The calcium metabolism is, however, primarily regulated by the Para Thyroid Hormone (PTH) and the calcium-sensing receptor (CASR). See, for example: Thakker R V: Disorders of the calcium-sensing receptor. Biochim Biophys Acta. 1998 Dec. 10;1448(2):166-70.
Further data and information about the behavior of the D metabolites in Th1 inflammation can be found in this inventor's collaborative publication: Waterhouse J C, Marshall T G, Fenter B, Mangin M, Blaney G: High levels of active 1,25-dihydroxyvitamin D despite low levels of the 25-hydroxyvitamin D precursor—Implications of dysregulated vitamin D for diagnosis and treatment of Chronic Disease. In Vitamin D: New Research. Volume 1. Edited by: Stoltz V D. New York: Nova Science Publishers; 2006. ISBN: 1-60021-000-7
Optimally Effective Dosing Regimes for Ligands Acting as GPCR and NR Antagonists
The subject compounds/ligands exhibit different affinities for different receptors. As the bloodstream concentration of each ligand is changed, they will have different effects on different receptors. An example chart showing such sensitivity variation for the ARBs is given in our paper “Marshall T G, Lee R E, Marshall F E: Common Angiotensin Receptor Blockers may directly modulate the immune system via VDR, PPAR and CCR2b. Theoretical Biology and Medical Modelling, not yet published” (draft attached).
Consequently the choice of ligands is important. One must employ ligands with good affinity for the receptors one wants to block, and with minimum affinity for those which are necessary for proper functioning of the body.
Additionally, the concentration of ligand in the bloodstream must be kept relatively constant, so as to help target those receptors which will enhance the health of the human body, and minimize the health of the pathogen(s).
This inventor has previously performed research on novel drug-dosing regimes in several diseases. In Diabetes, I explored increased efficacy and reduced side-effects from a continuous infusion of Insulin, and in Cryptorchidism and Infertility I explored increased efficacy of pulsatile dosing of the hormones LH-RH and Gn-RH.
During my Doctoral Research we also explored pulsatile administration of drugs, and my research group was able to cure Cryptorchidism and Infertility by using pulsatile injections of hormone, rather than using a continuous concentration: Keogh E J, MacKellar A, Mallal S A, Dunn A G, McColm S C, Somerville C P, Glatthaar C, Marshall T, Attikiouzel J: Treatment of cryptorchidism with pulsatile luteinizing hormone-releasing hormone (LH-RH). J Pediatr Surg. 1983 Jun; 18(3):282-3.
One of the best ways to administer a continuous concentration of any drug is by using an infusion pump, for example the transcutaneous infusion pump for Insulin I invented in 1982: Marshall T G, Mekhiel N, Jackman W S, Perlman K, Albisser A M: New microprocessor-based insulin controller. IEEE Trans Biomed Eng. 1983 Nov.;30(11):689-95.
Other methods of administering semi-continuous concentrations of any drug are transcutaneous patches, sub-dermal implanted ‘reservoirs’, controlled-release drug compounding formulations, controlled-release binders (such as polymers) and implanted infusion pumps.
For example, the administration of Statins and ARBs at intervals beyond about 8 hours causes them to lose efficacy, because the concentration in the blood stream drops below the level at which a complete blockade of the undesirable receptors is effected. Thus, the conventional dosing of the ARBs that the FDA approved for hypertension, 24 hourly, with “the amount of return on twice daily dosing—already a poor investment,” does not allow them to function effectively as pathogenic antagonists. They must be dosed much more frequently (preferably semi-continuously) so as to apply the maximum possible blockade to the Pathogenic Receptors in the inflamed tissue.
Special Considerations for Asserted Therapeutic or Pharmacological Utilities
With respect to MPEP 2107.03 “Special Considerations for Asserted Therapeutic or Pharmacological Utilities,” the utility of this methods patent has been established by “statistically relevant data documenting the activity of a compound or composition, arguments or reasoning, documentary evidence (e.g., articles in scientific journals), or any combination thereof.”
A sampling of peer-reviewed papers and conference presentations (from this inventor) sufficient to establish such utility, and which also document “actual evidence of success in treating humans” during the Phase 2 clinical studies conducted by this inventor, includes:                Marshall T G. What is the role of CWD bacteria during HIV infection? Invited Conference Presentation, ‘Recovering from Chronic Disease’, Jun. 17, 2006, Publisher: Autoimmunity Research Foundation (DVD transcript available)        Marshall T G: VDR Nuclear Receptor Competence is the Key to Recovery from Chronic Inflammatory and Autoimmune Disease. Abstract presentation, Days of molecular medicine, 2006. Copy available from URL http://autoimmunityresearch.org/karolinska-handout.pdf        Marshall T G, Lee R E, Marshall F E: Common Angiotensin Receptor Blockers may directly modulate the immune system via VDR, PPAR and CCR2b, Theoretical Biology and Medical Modelling,2006 Jan. 10;3(1):1. Available from URL http://www.tbiomed.com/content/3/1/1        Marshall T G: Molecular genomics offers new insight into the exact mechanism of action of common drugs—ARBs, Statins, and Corticosteroids. FDA CDER Visiting Professor presentation, FDA Biosciences Library, Accession QH447.M27 2006. Copy available from URL http://autoimmunityresearch.org/fda-visiting-professor-7mar06.ram        Marshall T G, Marshall F E: Sarcoidosis succumbs to antibiotics—implications for autoimmune disease. Autoimmunity Reviews, 2004; 3(4):295-3001.        Waterhouse J C, Marshall T G, Fenter B, Mangin M, Blaney G: High levels of active 1,25-dihydroxyvitamin D despite low levels of the 25-hydroxyvitamin D precursor—Implications of dysregulated vitamin D for diagnosis and treatment of Chronic Disease. In Vitamin D: New Research. Volume 1. Edited by: Stoltz V D. New York: Nova Science Publishers; 2006. ISBN: 1-60021-000-7        Marshall T G, Fenter B J, Marshall F E: Antibacterial Therapy Induces Remission in Sarcoidosis (in English). JOIMR 2005;3(1):2 Available from URL http://www.joimr.org/phorum/read.php?f=2&i=107&t=107        Marshall T G, Fenter B, Marshall F E: Antibacterial Therapy Induces Remission in Sarcoidosis. Herald MKDTS 2004g; Volume III: Release 1. (The Journal of the Interregional Clinical-Diagnostic Center, Kazan, published in Russian translation). Invited Paper. Special issue on Sarcoidosis. ISSN: 1726-6149        Marshall T G, Fenter B, Marshall F E: Putative Antibacterial Mechanisms for Angiotensin Receptor Blockers. JOIMR 2004;2(2):1.        Marshall T G , Marshall F E : Sarcoidosis succumbs to antibiotics—implications for autoimmune disease. Autoimmunity Reviews,2004; Supplement 2:55 (Abstracts of 4th International Congress on Autoimmunity)        Marshall T G : Bacterial Th1 Processes Seem Key to Chronic Lyme Remission. ILADS conference, October 2004, Rye Town, N.Y.        Marshall T G : How Borrelia Evades the Immune System, and How we Help it Kill This Th1 Bacterium. ‘30 th Anniversary of Lyme Disease’ conference, Farmington, Conn., May 7, 2005        Marshall T G , Mangin M, Marshall F E : Bacterial Th1 Processes Key to CFS/ME Remission. AACFS conference, Madison, Wis., October 2004        Marshall T G : Genomics, Molecular Medicine and Antibiotic Resistance. ‘Recovery From Chronic Disease’ conference, Chicago, Ill., Mar. 12, 2005.        